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STAMBP / STAM-binding protein · IHC design guide
Design Immunohistochemistry for STAMBP
Plan paraffin STAMBP IHC around nuclear and cytoplasmic staining (HPA tissue IHC). Breast and colon glandular cells show high staining (HPA tissue IHC); the catalog antibody lists 2–5 μg/ml for IHC (datasheet A05964-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
STAMBP Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | General nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05964-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | IHC staining has uncertain reliability versus RNA data (HPA tissue IHC) | |
| Regulation | Ubiquitously expressed; regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope impact unresolved (UniProt) |
Section 1
Recommended STAMBP IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A05964-1) with published STAMBP methods for human tissue and subcutaneous tumors (PMC11379802) and human LUAD tissue microarrays (PMC8190130).
| Sample | Paraffin-embedded rat testis tissue; fixative not specified (datasheet A05964-1) |
| Fixation | Image fixative and duration unreported (datasheet A05964-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A05964-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05964-1) |
| Primary antibody | Rabbit anti-STAMBP, 2-5 μg/ml (datasheet A05964-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05964-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05964-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | STAMBP-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet: A05964-1); consider EDTA pH 9.0 when optimizing LUAD sections (PMC8190130).
Section 2
What Is the Expected STAMBP Staining Pattern?
For paraffin-section IHC, expect nuclear and cytoplasmic staining in several cell types, including glandular cells in adrenal gland, appendix, breast, colon, duodenum, fallopian tube, and gallbladder, and glial cells in cerebral cortex (HPA tissue IHC: High; general nuclear and cytoplasmic expression). Treat this as a provisional pattern because tissue IHC reliability is Uncertain (HPA tissue IHC). STAMBP also associates with membranes and early endosomes but has no transmembrane segment (UniProt O95630: localization and topology).
| Nuclear and cytoplasmic signal in glandular cells of a listed high-staining tissue. | This fits the reported tissue pattern, especially when the signal is stronger than nearby background (HPA tissue IHC: general nuclear and cytoplasmic expression; High in glandular cells). Interpret intensity within the same run: the HPA label describes its observed staining, while its overall IHC reliability remains Uncertain (HPA tissue IHC). |
| Predominantly sharp staining in an unexpected compartment, with little nuclear or cytoplasmic signal. | Question the result before assigning a new IHC localization: HPA reports general nuclear and cytoplasmic tissue staining (HPA tissue IHC), while UniProt also lists membrane and early endosome localization (UniProt O95630). Check morphology and controls; an unusual pattern alone cannot establish either artifact or a genuine compartment-specific pool. |
| Strong staining in a cell population expected to stain weakly, such as esophageal squamous cells or adipocytes. | Investigate possible cross-reactivity or endogenous detection activity before calling it STAMBP: those cell populations are reported Low, while STAMBP is broadly expressed (HPA tissue IHC: Low in esophageal squamous cells and adipocytes; UniProt O95630: ubiquitous expression). Low is a comparison point, not a required absence of signal. |
| Diffuse colour across cells, extracellular spaces, and the section edge. | Treat a pattern without clear cell boundaries as background until controls support a cellular signal (general IHC practice). Compare a reagent-control section and inspect uneven counterstain or detection deposits (general IHC practice). The reported STAMBP pattern is cellular and generally nuclear and cytoplasmic (HPA tissue IHC). |
| No convincing signal in glandular cells of a listed high-staining tissue. | Review the run before interpreting the sample as STAMBP-negative: these cells were reported High by HPA, although the tissue IHC assessment is Uncertain (HPA tissue IHC). Check section quality, retrieval, antibody dilution, and detection controls as general IHC troubleshooting; no STAMBP-specific retrieval or fixation sensitivity is supplied. |
💡Expected STAMBP appearanceA plausible positive paraffin-section result is cellular nuclear and cytoplasmic staining in the listed glandular cells or cerebral-cortex glia, potentially strong in HPA's High groups, whereas colour distributed uniformly outside cells is suspicious background (HPA tissue IHC: pattern, cell types, and High levels; general IHC practice).
| Tissue IHC confidence | HPA calls its tissue staining Uncertain because antibody staining and RNA expression have medium consistency (HPA tissue IHC); interpret a match as support for the pattern, not independent proof of specificity. |
| Antibody-specific validation | HPA035800 is Uncertain for IHC and Supported for ICC; HPA035801 has Supported ICC status but no listed IHC status (HPA antibodies). Do not transfer an ICC validation label to paraffin-section IHC. |
| Cell type and comparison tissue | HPA reports High staining in specified glandular cells and cerebral-cortex glia, but Low staining in esophageal squamous cells and adipocytes (HPA tissue IHC). Score the stated cell population rather than a whole-section average. |
| Subcellular context | UniProt lists nucleus, cytoplasm, membrane, and early endosome, with no transmembrane segment (UniProt O95630: localization and topology). HPA tissue IHC reports general nuclear and cytoplasmic staining; its finer nucleoplasm, plasma-membrane, and cytosol assignments come from ICC-IF (HPA tissue IHC; HPA subcellular). |
| Isoforms and epitope scope | Two isoforms are listed (UniProt O95630: isoforms 1 and 2). Their presence alone does not establish that an antibody recognizes both; use the antibody's documented immunogen or epitope information when available (general IHC practice). No epitope location is supplied here. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed high-staining tissue appears blank. | The run may lack usable antigen or detection signal; the HPA High label is observational and its IHC reliability is Uncertain (HPA tissue IHC). | Check tissue preservation, retrieval, antibody dilution, and detection with routine controls (general IHC practice). No target-specific fixation or retrieval effect is established here. |
| Nuclear signal is absent but cytoplasmic signal remains. | The observed compartment balance may differ across cells or runs; HPA describes general nuclear and cytoplasmic tissue expression (HPA tissue IHC). | Compare matched cells and control sections, then document each compartment separately (general IHC practice); avoid declaring a new STAMBP localization from this slide alone. |
| Low-reference cells stain as strongly as the selected high-reference cells. | Cross-reactivity, endogenous chromogenic activity, or a run-wide background problem is possible (general IHC practice); HPA reports those cell groups as Low (HPA tissue IHC). | Compare reagent controls and inspect whether colour follows cellular structures (general IHC practice). Treat Low as relative staining, not a mandatory negative result (HPA tissue IHC). |
| Colour is diffuse or concentrated at section edges. | Uneven reagent coverage, nonspecific binding, or detection deposits can produce this appearance (general IHC practice). | Review blocking, washes, and detection controls, and score only interpretable cells (general IHC practice). The reported tissue pattern is cellular and generally nuclear and cytoplasmic (HPA tissue IHC). |
| Two antibodies give different IHC patterns. | Their validation information differs: HPA035800 has Uncertain IHC status, and no IHC status is listed for HPA035801 (HPA antibodies). | Keep antibody identities separate in the record and compare each result with cell type, compartment, and controls (HPA antibodies; HPA tissue IHC; general IHC practice). |
| IF/ICC: what localization should be checked? | HPA's ICC-IF summary places STAMBP mainly in the nucleoplasm, with additional plasma-membrane and cytosol localization (HPA subcellular: Supported). | Use the separate IF/ICC guide for that application; record nuclear, cytosolic, and membrane signal separately (HPA subcellular; general IF practice). |
Experiment evidence
IHC & IF images showing STAMBP staining
A05964-1 · IHCIHC analysis of STAMBP using anti-STAMBP antibody (A05964-1). STAMBP was detected in a paraffin-embedded section of rat testis tissue. Heat
A05964-1 · IHCIHC analysis of STAMBP using anti-STAMBP antibody (A05964-1). STAMBP was detected in a paraffin-embedded section of mouse testis tissue. Hea
A05964-1 · IFIF analysis of STAMBP using anti-STAMBP antibody (A05964-1). STAMBP was detected in an immunocytochemical section of PC3 cells. Enzyme antig
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000124356-STAMBP/tissue/cerebral+cortex#imid_8840988
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000124356-STAMBP/tissue/colon#imid_8840930
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000124356-STAMBP/tissue/liver#imid_8840915
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000124356-STAMBP/tissue/kidney#imid_8840924
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035800; green = STAMBP, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035800; green = STAMBP, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for STAMBP IHC & IF
🧪Run cerebral cortex first and score staining in glial cells, which HPA rates High (HPA: High in cerebral cortex glial cells). HPA detects STAMBP in all 45 scored tissues, so there is no supported negative tissue or validated negative cell population within the slide; no-primary and isotype controls establish background, and cells lacking specific staining should show counterstain only (HPA: detected in all 45 scored tissues; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: STAMBP is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show STAMBP in A-431, U-251MG, U2OS, LHCN-M2, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an isotype control matched to the rabbit primary antibody, using the same detection conditions (selected-SKU IHC caption: rabbit primary and goat anti-rabbit secondary; standard IHC practice). Use STAMBP knockout tissue or cells as a biological negative, and block endogenous peroxidase before HRP–DAB detection in cerebral cortex sections (selected-SKU IHC caption: HRP–DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU IHC caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; the supplied evidence does not establish whether frozen sections or IF are easier for STAMBP (selected-SKU IHC caption: EDTA retrieval; HPA: ICC-IF localization evidence). In cerebral cortex, lipofuscin pigment or autofluorescence can complicate interpretation, so assess staining against the matched controls and cellular morphology (standard IHC/IF practice).
HPA tissue IHC evidence for STAMBP
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: STAMBP is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced STAMBP IHC Tips
Troubleshoot STAMBP staining in paraffin sections by checking retrieval, compartment pattern, cell identity and scoring against the available tissue evidence.
Which retrieval condition should I start with for STAMBP in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for this catalog antibody (datasheet A05964-1). The selected paraffin-section image used that condition before detecting STAMBP in rat testis (caption A05964-1). If staining is weak, compare retrieval durations on adjacent sections while keeping the antibody concentration and detection conditions constant (standard IHC practice). Use an unstained control section to check whether longer heating increases tissue damage or nonspecific color (standard IHC practice). Record the heating device, temperature and duration, since the supplied caption does not specify them (caption A05964-1).
Could fixation explain weak or uneven STAMBP staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A05964-1). Record the actual fixative and fixation duration for each specimen, then compare sections processed under matched retrieval and staining conditions (standard IHC practice). If archival blocks vary in handling, assess morphology and staining in a consistently processed control alongside each run (standard IHC practice). Keep section thickness, retrieval and development time consistent before attributing an intensity difference to fixation (standard IHC practice). The image documents staining after EDTA pH 8.0 retrieval, but cannot establish how STAMBP responds to any particular fixative (caption A05964-1).
What cellular staining pattern is plausible for STAMBP?
Assess nuclear and cytoplasmic staining separately: tissue IHC reports both compartments, with uncertain staining reliability (HPA tissue IHC). Nucleoplasmic signal is supported by cellular imaging, with additional plasma-membrane and cytosolic localisation (HPA subcellular). Early-endosome localisation is also reported, so a punctate cytoplasmic component can be biologically plausible (UniProt O95630). STAMBP has no transmembrane segment; do not interpret a membrane outline alone as proof that the protein spans the membrane (UniProt O95630 topology). Compare compartment patterns within intact cells and across matched control sections before scoring diffuse pigment as STAMBP (standard IHC practice).
How do isoforms and epitope placement affect a negative IHC result?
STAMBP has 2 annotated isoforms, but the supplied antibody evidence does not map its epitope to either one (UniProt O95630; caption A05964-1). The MPN domain spans residues 257–388, and several phosphorylation sites are annotated elsewhere in the sequence (UniProt O95630). Those annotations do not establish that fixation or retrieval masks this antibody’s epitope (UniProt O95630; caption A05964-1). If staining is absent, first check the same-run positive control, retrieval and detection steps before proposing isoform-specific loss (standard IHC practice). Report isoform recognition as unknown unless epitope mapping or direct isoform testing becomes available (caption A05964-1).
How should I use IF to investigate the IHC staining pattern?
Use IF as a separate localisation check; the catalog antibody’s supplied tissue image documents chromogenic IHC, not an IF protocol (caption A05964-1). For multiplexing, pair STAMBP with an epithelial marker when examining glandular cells, a reported high-staining population in several tissues (HPA tissue IHC; standard IF practice). Choose a red or far-red fluorophore after checking tissue autofluorescence in each channel, and include single-label controls for bleed-through (standard IF practice). Because STAMBP lacks a transmembrane segment and can localise to the nucleoplasm and cytosol, use controlled permeabilisation when assessing intracellular signal (UniProt O95630 topology; HPA subcellular; standard IF practice). The antibody epitope’s accessibility remains unknown (caption A05964-1).
How can I reduce diffuse DAB staining without losing STAMBP signal?
The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase-based DAB detection (caption A05964-1). Reproduce those stated conditions first, then compare adjacent sections with and without primary antibody to locate detection-related background (caption A05964-1; standard IHC practice). Include an endogenous-peroxidase blocking step and control DAB development time, treating both as general chromogenic IHC practices (standard IHC practice). If diffuse color persists, titrate primary antibody around the documented concentration while keeping retrieval constant (caption A05964-1; standard IHC practice). Check whether pigment also appears outside intact cells before assigning it to STAMBP (standard IHC practice).
What is a defensible way to score STAMBP IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear and cytoplasmic staining separately, because both compartments are reported for STAMBP (HPA tissue IHC). For each compartment, record the percentage of positive cells and an intensity-based H-score, using identical thresholds across matched sections (standard IHC practice). Normalise positive-cell counts to the number of evaluable cells in the same annotated region; use density per mm² only when region area is measured (standard IHC practice). Exclude folds, necrosis and section edges by a rule set before viewing group labels (standard IHC practice). Report the antibody, retrieval condition and uncertain HPA tissue-staining reliability alongside the scores (caption A05964-1; HPA tissue IHC).
How can I distinguish convincing STAMBP staining from artefact?
Convincing signal should occur within intact cells in plausible nuclear or cytoplasmic compartments; STAMBP also has reported early-endosome localisation (HPA tissue IHC; UniProt O95630). Compare cell identity with the tissue context: high glandular-cell staining is reported in several tissues, while squamous epithelial cells in esophagus and adipocytes have low reported staining (HPA tissue IHC). Reject signal confined to section edges, folds or necrotic areas after checking the matched morphology (standard IHC practice). A no-primary control helps identify endogenous-enzyme or detection-related DAB color (standard IHC practice). Treat discordant patterns cautiously because HPA rates its tissue IHC reliability uncertain (HPA tissue IHC).
Boster reagents
Best STAMBP / STAM-binding protein IHC Antibodies
A05964-1 has IHC images from human, mouse and rat paraffin sections and an IF image from PC3 cells (catalog: IHC and IF image captions).
Real IHC data
Anti-STAMBP Antibody ®
Cat # A05964-1
A05964-1 is shown in IHC on rat and mouse testis, human ovarian cancer tissue and human spleen (catalog: IHC image captions). The same SKU is shown in IF on PC3 cells (catalog: IF image caption).
Which to pick: For tissue IHC, choose A05964-1 for paraffin sections; its images use heat retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml (catalog: IHC image captions). For IF/ICC, A05964-1 is listed at 5 μg/ml and shown on PC3 cells (catalog: applications, IF dilution and IF image caption). For cross-species work, A05964-1 lists human, monkey, mouse and rat reactivity, with IHC images for human, mouse and rat; it is rabbit hosted, while clonality and the paraffin sections’ fixative are unreported (catalog: reactivity, host and clone; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95630 (STABP_HUMAN, STAM-binding protein).
- Human Protein Atlas. STAMBP tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. STAMBP subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane and cytosol..
- Human Protein Atlas. STAMBP antibody validation summary (2 antibodies).
- The deubiquitinating enzyme STAMBP is a newly discovered driver of triple-negative breast cancer progression that maintains RAI14 protein stability. Experimental & molecular medicine 2022 — PMC9723177.
- Identification of STAM-binding protein as a target for the treatment of gemcitabine resistance pancreatic cancer in a nutrient-poor microenvironment. Cell death & disease 2024 — PMC11379802.
- STAMBP promotes lung adenocarcinoma metastasis by regulating the EGFR/MAPK signaling pathway. Neoplasia (New York, N.Y.) 2021 — PMC8190130.
- PubMed PMID:10383417 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for STAMBP (STAM-binding protein). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05964-1). Validated IHC antibody: A05964-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat testis tissue; fixative not specified (datasheet A05964-1); 2. Fixation: Image fixative and duration unreported (datasheet A05964-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A05964-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05964-1); 8. Primary antibody: Rabbit anti-STAMBP, 2-5 μg/ml (datasheet A05964-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05964-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: IHC staining has uncertain reliability versus RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope impact unresolved (UniProt); Regulation: Ubiquitously expressed; regulation unreported (UniProt). Evidence: UniProt O95630, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05964-1, PMC11379802, PMC8190130. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/STAMBP/immunohistochemistry-ihc-guide.